ZNK1 senses zinc and degrades zinc transporter mRNA in trypanosomes

Abstract

Like other cells, parasitic and other trypanosomatids sense and regulate Zn2+ transport, but the mechanisms involved remained unknown. Here we identify a trypanosome RNA-binding protein which specifically eliminates ZIP3 Zn2+-transporter mRNA in Zn2+-replete conditions. We first demonstrated that Trypanosoma brucei ZIP3 mRNA abundance is subject to 3-untranslated region (3-UTR) and Zn2+-dependent negative control. A genome-wide RNA interference library screen, using a reporter associated with the ZIP3 3-UTR, identified Tb927.11.9510 as a candidate Zn2+-sensor. We name this protein Zinc Nuclear Knuckles 1 (ZNK1) since it localises to the nucleus and contains several Zn2+-knuckle motifs. ZNK1 is conserved among trypanosomatids, and a PIN-domain suggests a ribonuclease-based mechanism. We validate ZNK1 as a ZIP3 3-UTR dependent negative regulator and identify a GU-repeat motif in the ZIP3 3-UTR that is predictive of negative control by ZNK1. We use Cas9-editing to knockout ZNK1, and RNA-seq to assess the consequences, revealing highly specific accumulation of ZIP3 transcripts in znk1-null cells. We conclude that ZNK1 senses Zn2+-abundance and eliminates ZIP3 mRNA in a Zn2+-dependent manner. We suggest that trypanosomatid ZNK1 is an RNA-specific zinc finger nuclease that binds ZIP3 3-UTRs and degrades ZIP3 mRNA only when the tandem sensor modules are coordinated with Zn2+. Key pointsO_LITrypanosome Zinc Nuclear Knuckles 1 (ZNK1) is a zinc-sensor that eliminates zinc transporter mRNA. C_LIO_LIZNK1 negative control operates via the transporter mRNA 3-untranslated region. C_LIO_LIThe findings indicate that trypanosomatid ZNK1 is a conserved RNA-specific zinc finger nuclease. C_LI